Myeloid Activation Assay Staining Protocol

This protocol is for reference purposes only. It may be necessary to adapt the method for specific research needs.

Materials

IOTest Myeloid Activation CD169-PE/HLA-DR-APC/CD64-PB Antibody Cocktail, Part Number C63854

MATERIALS REQUIRED, BUT NOT PROVIDED

Whole blood collection tubes with EDTA or Heparin anticoagulant

12 x 75 mm test tubes (Part Number 2523749)

Pipettors and pipet tips capable of delivering 10 and 500 µL

VersaLyse Solution (Part Number IM3648)

IOTest 3 Fixative Solution, 10X concentrate (Part Number IM3515)

VersaLyse/Fixative:
Dilute IOTest 3 Fixative Solution 1:220 in VersaLyse Solution

Activators used for positive control

Flow cytometer

Staining Procedure

  1. Add 5 to 25 μL of whole blood to test tube.
  2. Add 10 μL of the IOTest CD169-PE/ HLA-DR-APC/ CD64-PB cocktail.
  3. Add 500 μL of VersaLyse/Fixative.
  4. Incubate 10 minutes.
  5. Acquire data with flow cytometer.

Note: For heparin anti-coagulated blood, stained samples can be acquired for up to 6 hours. For EDTA anti-coagulated blood, stained samples can be acquired for up to 3 hours.

See Example Data